Multiparameter label-free flow cytometry using multiplex coherent anti-Stokes Raman scattering (MCARS) with biological applications

Charles H. Camp Jr.; Siva Yegnanarayanan; Ali A. Eftekhar; Ali Adibi

doi:10.1117/12.874998

11 February 2011 Multiparameter label-free flow cytometry using multiplex coherent anti-Stokes Raman scattering (MCARS) with biological applications

Charles H. Camp Jr., Siva Yegnanarayanan, Ali A. Eftekhar, Ali Adibi

Author Affiliations +

Proceedings Volume 7903, Multiphoton Microscopy in the Biomedical Sciences XI; 79031G (2011) https://doi.org/10.1117/12.874998
Event: SPIE BiOS, 2011, San Francisco, California, United States

Abstract

Flow cytometry is an invaluable analytical tool that provides statistical information about sample populations. By optically probing samples at high-speed, flow cytometers retrieve information about basic morphology and composition. The optical elastic scatter is indicative of sample size and granularity while molecular information requires the addition of fluorescent labels, which have limitations such as spectral overlap, non-specific binding, and cellular toxicity. In this work, we present the first multiparameter label-free flow cytometer that observes the elastically forward-scattered light (FSC) and probes the intrinsic Raman vibrations of passing samples using multiplex coherent anti-Stokes Raman scattering (MCARS). MCARS, as a broadband technique, probes a large region of the Raman spectrum; thus, leading to rich molecularly-sensitive information. Additionally, we present the first experimental investigations of two biological systems using the multiparameter flow cytometer: Saccharomyces cerevisiae, a yeast often used as a model system for eukaryotic organisms, and Phaeodactylum tricornutum, a diatom under investigation for biofuel production.

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Charles H. Camp Jr., Siva Yegnanarayanan, Ali A. Eftekhar, and Ali Adibi "Multiparameter label-free flow cytometry using multiplex coherent anti-Stokes Raman scattering (MCARS) with biological applications", Proc. SPIE 7903, Multiphoton Microscopy in the Biomedical Sciences XI, 79031G (11 February 2011); https://doi.org/10.1117/12.874998

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KEYWORDS

Yeast

Raman spectroscopy

Flow cytometry

Nitrogen

Spectroscopy

Organisms

Microfluidics

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